Rho D, Hodgson J, Thiboutot S, Ampleman G, Hawari J
Biotechnology Research Institute, National Research Council (NRC) Canada, 6100 Royalmount Avenue, Montréal, QC, H4P 2R2, Canada.
Biotechnol Bioeng. 2001 May 20;73(4):271-81. doi: 10.1002/bit.1060.
The cometabolic transformation of 2,4,6-trinitrotoluene (TNT) by an immobilized Phanerochaete chrysosporium culture was investigated under different TNT and/or glycerol feeding conditions in a 5-L reactor. In the fed-batch feeding mode, as a result of four spiking events at an average feeding rate of 20 mg TNT L(-1) d(-1) and 250 mg glycerol L(-1) d(-1), the initial TNT transformation rate and the glycerol uptake rate of the 7-day-old immobilized cell culture were 2.41 mg L(-1) h(-1) and 16.6 mg L(-1) h(-1), respectively. Thereafter, the TNT fed into the reactor depicted a negative effect on the cell physiology of P. chrysosporium, i.e., both rates decreased constantly. At 32 mg TNT L(-1) d(-1) feeding rate, also in the presence of glycerol (200 mg L(-1) d(-1)), this effect on the fungal cell metabolism was even more significant. When TNT was fed alone at 3.7 mg L(-1) d(-1), it showed an initial 0.75 mg L(-1) h(-1) rate of TNT transformation, i.e., one-third the initial level observed in the presence of glycerol. In contrast, in the continuous feeding mode (dilution rate, D = 0.11 d(-1)), at 5.5 mg TNT L(-1) d(-1) and 220 mg glycerol L(-1) d(-1), the immobilized cell culture exhibited a constant TNT transformation rate for cultivation periods of 50 and 61 days, under uncontrolled and controlled pH conditions, respectively. Thereafter, during the latter experiment, 100% TNT biotransformation was achieved at 1,100 mg L(-1) d(-1) glycerol feeding rate. Immobilized cells (115-day-old), sampled from a continuous TNT feeding experiment, mineralized [(14)C]-TNT to a level of 15.3% following a 41-day incubation period in a microcosm.
在5升反应器中，研究了在不同的三硝基甲苯（TNT）和/或甘油进料条件下，固定化的黄孢原毛平革菌培养物对TNT的共代谢转化。在分批补料进料模式下，由于以20毫克TNT L⁻¹ d⁻¹和250毫克甘油L⁻¹ d⁻¹的平均进料速率进行了四次加样事件，7日龄固定化细胞培养物的初始TNT转化率和甘油摄取率分别为2.41毫克L⁻¹ h⁻¹和16.6毫克L⁻¹ h⁻¹。此后，进料到反应器中的TNT对黄孢原毛平革菌的细胞生理学产生了负面影响，即两种速率都持续下降。在32毫克TNT L⁻¹ d⁻¹的进料速率下，同样在存在甘油（200毫克L⁻¹ d⁻¹）的情况下，这种对真菌细胞代谢的影响更为显著。当单独以3.7毫克L⁻¹ d⁻¹进料TNT时，其显示出初始0.75毫克L⁻¹ h⁻¹的TNT转化速率，即存在甘油时观察到的初始水平的三分之一。相比之下，在连续进料模式（稀释率，D = 0.11 d⁻¹）下，在5.5毫克TNT L⁻¹ d⁻¹和220毫克甘油L⁻¹ d⁻¹的条件下，固定化细胞培养物在分别不受控和受控pH条件下培养50天和61天期间表现出恒定的TNT转化率。此后，在后者的实验中，在1100毫克L⁻¹ d⁻¹的甘油进料速率下实现了100%的TNT生物转化。从连续TNT进料实验中取样的固定化细胞（115日龄），在微观世界中经过41天的孵育期后，将[¹⁴C]-TNT矿化至15.3%的水平。
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